JP5047627B2 - Pyridinium and quinolinium derivatives - Google Patents

Abstract

The invention provides compounds of formula I blocking phosphorylcholine biosynthesis by means of the selective blocking of the choline kinase enzyme in tumor cells or in cells affected by parasitic infection and therefore being applicable in the treatment of tumors and parasitic diseases or diseases produced by viruses and fungi in animals, including human beings; as well as to a method for preparing the compounds of the invention and certain intermediates of said method.

Description

Field of Invention

  The present invention generally relates to compounds that block phosphorylcholine biosynthesis by means of selective blocking of the choline kinase enzyme in tumor cells or cells affected by infection with parasites, and thus tumors and parasitic diseases in animals, including humans. Alternatively, it relates to compounds applicable to the treatment of diseases caused by viruses, bacteria and fungi, as well as methods for the preparation of the compounds of the invention and certain intermediates of the methods.

Background of the Invention

Choline kinase is the first enzyme in the Kennedy or phosphatidylcholine (PC) synthesis pathway, and phosphorylcholine (PCho) is phosphorylated by phosphorylating choline using adenosine 5 ′ triphosphate (ATP) as the phosphate donor. Kent, C. Prog. Lipid Res., 29, 87-105 (1990); Kennedy, EP Fed. Proc., 20, 934-940 (1961)]. Ras genes form a family called oncogenes and are widely studied because they are activated in 25-30% of all human tumors, and in some others 90% [Bos, JL. Cancer Res 49, 4682-4689 (1989); Kiaris, H., Spandidos, DA Int. J. Oncol., 413-421 (1995)]. Ras proteins play an important role in intracellular signaling due to their involvement in the regulation of cell proliferation, terminal differentiation and senescence [Abdellatif, M., MacLellan, WR; Schneider, MDJ Biol. Chem., 269, 15423-15426 (1994); Wiesmuller, L., Wittinghofer, F. Cell Signal., 6, 247-267 (1994); Barbacid, M. Eur. J. Clin. Invest., 20, 225-235 (1990) Hahn & Weinberg Nat. Rev. Cancer, 2: 331 (2002); Wright & Shay Nat. Biotech, 20: 682 (2002); Drayton & Peters Curr. Op. Gen. Dev, 12:98 (2002)]. Malignant transformation mediated by various oncogenes, especially the ras oncogene, induces high levels of choline kinase activity, resulting in abnormal increases in the intracellular levels of the product PCho [Lacal et al ., Nature 330, 269-272 (1987); Lacal JC Mol. Cell. Biol. 10, 333-340 (1990); Teegarden, D., Taparowsky, EJ, Kent, CJ Biol. Chem. 265, 6042-6047 (1990); Ratnam, S .; Kent, C. Arch. Biochem. Biophys. 323, 313-322 (1995); Ramirez de Molina, A., Rodriguez-Gonzalez, A., Penalva, V., Lucas, L ., Lacal, JC Biochem. Biophys. Res. Commun. 285, 873-879 (2001); Ramirez de Molina, A., Penalva, V .; Lucas, L., Lacal, JC Oncogene 21, 937-946 (2002) )]. Supplementary studies using nuclear magnetic resonance (NMR) technology have shown high levels of PCho in normal tissues, especially human tumor tissues including breast cancer, colon cancer, lung cancer and prostate cancer The facts also support ChoK's role in human tumor formation [Ruiz-Cabello, J., Cohen, JS NMR Biomed. 5, 226-233 (1992); de Certaines, JD, Larsen, VA, Podo, F., Carpinelli, G., Briot, 0., Henriksen, 0. NMR Biomed. 6, 345-365 (1993); Smith, TAD, Bush, C., Jameson, C., Titley, JC, Leach, MO, Wilman, DEV, McCready, VR NMR Biomed. 6, 318-323 (1993)]. ras is one of the most studied oncogenes in human carcinogenesis, and ChoK inhibition has been shown to be a novel and efficient anti-tumor strategy in cells malignantly transformed by oncogenes It is general knowledge [Cuadrado, A., Carnero, A., Dolfi, F., Jimenez, B., Lacal, JC Oncogene, 8, 2959-2968 (1993); Jimenez, B., del Peso , L., Montaner, S., Esteve, P. Lacal, JCJ Cell Biochem., 57, 141-149 (1995); Hernandez-Alcoceba, R., Saniger, L., Campos, J., Nunez, MC, Khaless, F., Gallo, MA, Espinosa, A., Lacal, JC Oncogene, 15, 2289-2301 (1997)]. These first findings were subsequently estimated in vivo in nude mice [Hernandez-Alcoceba, R., Fernandez, F., Lacal, JC Cancer Res. 59, 3112-3118 (1999)]. Studies on ChoK inhibitors have identified hemicolinium-3 (HC-3) as a relatively potent and selective blocker [Cuadrado A., Carnero A., Dolfi F., Jimenez B. and Lacal JC Oncogene 8, 2959-2968 (1993); Jimenez B., del Peso L., Montaner S., Esteve P. and Lacal JCJ cell Biochem. 57, 141-149 (1995); Hernandez-Alcoceba, R., Saniger, L., Campos, J., Nunez, MC, Khaless, F., Gallo, MA, Espinosa, A., Lacal, JC Oncogene, 15, 2289-2301 (1997)]. This choline homologue with a biphenyl structure has been used to design new antitumor drugs. Since HC-3 is a powerful respiratory palsy, it is not a good candidate for clinical use. The synthesis of some derivatives is based on structural modifications of HC-3 that improve ChoK inhibitory activity and suppress its toxic effects. The inhibitory effect produced by bis quaternized symmetrical compounds during growth has been linked to the ability to induce whole-cell PCho production [Hernandez-Alcoceba, R., Saniger, L., Campos, J., Nunez, MC, Khaless, F., Gallo, MA, Espinosa, A., Lacal, JC Oncogene, 15, 2289-2301 (1997) and ES 2 117 950]. When 1,2-ethylene-p- (bibenzyldimethyl-diyl) residue is used as a spacer between two cationic pyridinium heads substituted at the 4-position [Campos, J., Nunez, MC, Rodriguez , V., Gallo, MA, Espinosa, A. Bioorg. & Med. Chem. Lett. 10, 767-770 (2000)], their structure has been evaluated by their ability to inhibit isolated ChoK. (Ex vivo conditions) [Lacal JC IDrugs 4: 419 426 (2001)]. This 4-NR ₂ group has made a significant contribution, and due to the delocalization of the positive charge, it has been proposed that the role of this group is electrical [Campos, J., Nunez, MC, Rodriguez, V., Gallo, MA, Espinosa, A. Bioorg. & Med. Chem. Lett. 10, 767-770 (2000)]. Increased ChoK activity in various human breast cancers has been published [Ramirez de Molina, A., Gutierrez, R., Ramos, MA, Silva, JM, Silva, J., Sanchez, JJ, Bonilla, F., Lacal , JC Oncogen 21, 4317-4322 (2002)]. Recently, it has been reported that ChoK changes frequently occur in several human tumors such as lung cancer, colorectal cancer and prostate cancer [Ramirez de Molina, A., Rodriguez-Gonzalez, A., Gutierrez, R Martinez-Pinero, L., Sanchez, JJ, Bonilla, F., Resell, R., Lacal, JC Biochem. Biophys. Res. Commun. 296, 580-583 (2002)].

  However, the current state of the art, in particular the bis-quaternized pyridinium derivatives described in patent ES 2 117 950, exhibits a high level of toxicity and their therapeutic application is limited.

  Thus, there is a need in the present state of the art to develop compounds that have activity to block phosphorylcholine biosynthesis in tumor cells or in processes caused by parasitic, viral, bacterial or fungal infections while at the same time having low levels of toxicity. There is.

  The authors of the present invention have found that after extensive research, certain modifications in the state of the art, in particular the structure of the compounds described in patent ES 2 117 950, are unexpected and therefore surprising. It has been found that the level of toxicity of such compounds in the state of the art is significantly reduced.

BRIEF DESCRIPTION OF THE INVENTION

を有する一系列の化合物を提供し、この構造は、スペーサーにより連結された２つのＮ−アリールアミノピリジニウム基を有することを特徴とする。腫瘍細胞において、または可能性としては寄生虫、ウイルス、細菌または真菌感染によってもたらされるプロセスにおいてコリンキナーゼ酵素を選択的に遮断する手段により、ホスホリルコリン生合成の遮断薬として作用することに加え、この系列の化合物は低い毒性を有する。
第二の目的では、本発明は、薬剤における式Ｉの化合物の使用を提供する。 Thus, the present invention has as its first object the formula I:

A series of compounds is provided, the structure being characterized by having two N-arylaminopyridinium groups linked by a spacer. In addition to acting as a blocker of phosphorylcholine biosynthesis by means of selectively blocking choline kinase enzymes in tumor cells or possibly in processes caused by parasitic, viral, bacterial or fungal infections, this family This compound has low toxicity.
In a second object, the present invention provides the use of a compound of formula I in medicine.

A further object of the invention consists in providing a pharmaceutical formulation comprising at least one compound of formula I.
The present invention, in another object, provides a method for preparing a compound of formula I.

の化合物を提供する。 Finally, the present invention relates to the formula VII involved as starting compound in the process for the preparation of the compound of formula I:

Of the compound.

Detailed Description of the Invention

［式中、
Ｑ⁻は、医薬上好適な有機酸または無機酸の共役塩基を表し；
Ｒ_１およびＲ’_１は、互いに独立に、Ｈ、および場合によってトリフルオロメチル、ヒドロキシルまたはアルコキシルにより置換されていてもよいＣ_１−６アルキルからなる群から選択される基を表し；
Ｒ_２およびＲ’_２は、互いに独立に、場合によってハロゲン、トリフルオロメチル、ヒドロキシル、Ｃ_１−６アルキル、アミノまたはアルコキシルにより置換されていてもよいアリール基を表し；
Ｒ_３およびＲ’_３は、互いに独立に、Ｈ、ハロゲン、トリフルオロメチル、ヒドロキシル、アミノ、アルコキシル、および場合によってトリフルオロメチル、ヒドロキシル、アミノまたはアルコキシルにより置換されていてもよいＣ_１−６アルキルからなる群から選択される基か、あるいはそれぞれＲ_４およびＲ’_４とともに、互いに独立に、場合によってハロゲン、トリフルオロメチル、ヒドロキシル、Ｃ_１−６アルキル、アミノまたはアルコキシルにより置換されていてもよい−ＣＨ＝ＣＨ−ＣＨ＝ＣＨ−基のいずれかを表し；
Ｒ_４およびＲ’_４は、互いに独立に、Ｈ、および場合によってハロゲン、トリフルオロメチル、ヒドロキシル、アミノまたはアルコキシルにより置換されていてもよいＣ_１−６アルキルからなる群から選択される基か、あるいはそれぞれＲ_３およびＲ’_３とともに、互いに独立に、場合によってハロゲン、トリフルオロメチル、ヒドロキシル、Ｃ_１−６アルキル、アミノまたはアルコキシルにより置換されていてもよい−ＣＨ＝ＣＨ−ＣＨ＝ＣＨ−基のいずれかを表し；かつ、
Ａは、スペーサー基を表す］
に対応する一系列の化合物を提供する。 In its first object, the present invention provides a compound of general formula I:

[Where:
Q ⁻ represents a conjugated base of a pharmaceutically suitable organic or inorganic acid;
R ₁ and R ′ ₁ represent, independently of each other, a group selected from the group consisting of H and optionally C _1-6 alkyl optionally substituted by trifluoromethyl, hydroxyl or alkoxyl;
R ₂ and R ′ ₂ independently of one another represent an aryl group optionally substituted by halogen, trifluoromethyl, hydroxyl, C _1-6 alkyl, amino or alkoxyl;
R ₃ and R ′ ₃ are, independently of each other, H, halogen, trifluoromethyl, hydroxyl, amino, alkoxyl, and optionally C _1-6 alkyl optionally substituted by trifluoromethyl, hydroxyl, amino or alkoxyl A group selected from the group consisting of or independently of each other, together with R ₄ and R ′ ₄ , optionally substituted by halogen, trifluoromethyl, hydroxyl, C _1-6 alkyl, amino or alkoxyl Represents any of the groups -CH = CH-CH = CH-;
R ₄ and R ′ ₄ are, independently of each other, H and a group selected from the group consisting of C _1-6 alkyl optionally substituted by halogen, trifluoromethyl, hydroxyl, amino or alkoxyl; Or each independently with R ₃ and R ′ ₃ independently of each other —CH═CH—CH═CH— group optionally substituted by halogen, trifluoromethyl, hydroxyl, C _1-6 alkyl, amino or alkoxyl Any one of; and
A represents a spacer group]
A series of compounds corresponding to is provided.

  In addition to acting as a blocker of phosphorylcholine biosynthesis by means of selectively blocking the choline kinase enzyme in tumor cells or in cells affected by parasitic infections, compounds belonging to this family have been identified in the state of the art. It is characterized by having a lower level of toxicity than that of known structural compounds. This feature of the compounds of the invention is demonstrated in the examples given below.

式中、ｍ、ｎおよびｐは次の値：ｍ＝０、１；ｎ＝０、１〜１０；ｐ＝０、１を持ち得る整数を表す（ただし、ｍ、ｎおよびｐは同時に０の値をとることはない）。

In the context of the present invention, a spacer group “A” is understood as any divalent organic structure that acts as a linkage between two pyridinium groups present in the structure defined by formula I. In a particular embodiment of the invention, the spacer A has a structure according to one of the formulas II, III, IV, V and VI. These formulas indicate groups in which the terminal straight line-indicates a bond and not a methyl group.

Where m, n and p represent integers which can have the following values: m = 0, 1; n = 0, 1-10; p = 0, 1 (where m, n and p are simultaneously 0 Never take a value).

According to the invention, the groups R ₁ and R ′ ₁ , R ₂ and R ′ ₂ , and R ₃ and R ₄ , R ′ ₃ and R ′ ₄ may represent different groups or the same group, Asymmetric or symmetric compound.

In a particular embodiment of the invention, the radicals R ₂ and R ′ ₂ are, independently of one another, a phenyl group optionally substituted by halogen, trifluoromethyl, hydroxyl, C _1-6 alkyl, amino and alkoxyl. Represents. In another particular embodiment of the invention, the groups R ₁ and R ′ ₁ represent a methyl group, while the groups R ₂ and R ′ ₂ are independently of one another optionally substituted by one or more halogen substituents Represents an optionally substituted phenyl group. In a third particular embodiment, the groups R ₃ and R ₄ and the groups R ′ ₃ and R ′ ₄ are independently of each other but optionally together substituted by one or more halogen substituents -CH = CH-CH = CH- group which may be made.

Preferred compounds of the invention are shown in Table I below.

Finally, in a preferred embodiment of the invention, the pharmaceutically suitable organic or inorganic acid conjugate base Q represents Br (bromide) or F ₆ P (hexafluorophosphate).
The compounds of the present invention have a selective effect on the signaling pathways required to malignantly transform certain oncogenes, which do not act on normal cells at the same intensity and thus have great potency in anti-tumor treatment. Leave enough room.

  On the other hand, in the biological assay performed by the authors of the present invention, some parasites such as Plasmodium falciparum or Trypanosoma cruzi, some viruses such as adenovirus, pneumonia chain Bacteria such as Streptococcus pneumoniae and fungi such as Candida albicans require a metabolic pathway for phosphatidylcholine synthesis via choline kinases to complete their infection cycle in humans and animals. Is known to extend this type of activity to antiviral, antiparasitic, or antifungal activity. In this regard, the background in the literature is the role of ChoK in intracellular metabolism at specific nucleosides in Hep-G2 cells [Martin, LT; Faraj, A .; Schinazi, RF; Gosselin, G .; Mathe, C .; Imbach , J.-L .; Sommadossi, J.-P. Biochemical Pharmacology, 53, 75-87 (1997)], the use of ChoK as an enzyme marker in parasitic diseases [Wunderlich, F .; Helwig, M .; Schillinger , G .; Vial, H .; Philippot, J .; Speth, V. Molecular and Biochemical Parasitology, 23, 103-115 (1987); Ancelin, ML; Vial, HJ Biochimica et Biophysica Acta (BBA)-Lipids and Lipid Metabolism, 875, 52-58 (1986)] and viruses [Balakivera L., Schoen G., Thouvenin E., Chroboczek JJ Virol. 77: 4858-4866 (2003)], bacteria [Whiting GC, Gillespie SH. FEMS Microbiol Lett. 138: 141-145 (1996)] and fungi [Mago N, Khuller GK. J Med Vet Mycol. 28: 355-362 (1990)]); Mago N, Khuller GK. J Med Vet Mycol. 28: 355-362 (1990)] for biosynthesis of important phospholipids This confirms the involvement of ChoK. All these studies support that ChoK inhibition can have important therapeutic outcomes in the healing of the diseases described above.

  Thus, in a second object, the present invention provides the use of a compound of formula I in medicine. Specifically, the compounds of formula I are claimed for their use in medicine. In certain embodiments, the present invention provides compounds of formula I for the treatment of cancer, preferably breast cancer, lung cancer, colorectal cancer and pancreatic cancer. In another particular embodiment, the present invention provides for the treatment of viral diseases, particularly those caused by adenoviruses; and for the treatment of diseases caused by antiparasitic treatments, preferably plasmodium or trypanosoma; antibacterial treatments, preferably Provided are compounds of formula I for the treatment of diseases caused by streptococci; and the treatment of diseases caused by antifungal treatment, preferably Candida.

  On the other hand, the use of a compound of formula I in the manufacture of a medicament is claimed. In certain embodiments, the compounds of formula I are used in the manufacture of a medicament for cancer, preferably breast cancer, lung cancer, colorectal cancer or pancreatic cancer. In another particular embodiment, the compounds of formula I are used in the treatment of viral diseases, preferably in the manufacture of a medicament for diseases caused by adenoviruses; and in antiparasitic treatments, preferably caused by plasmodium or trypanosoma. For the manufacture of a medicament for the treatment of illnesses; for the treatment of bacterial diseases, preferably for the manufacture of diseases for diseases caused by streptococci; and for the treatment of fungal diseases, preferably for diseases caused by Candida Used in the manufacture of drugs for.

  In its third object, the present invention provides a pharmaceutical formulation comprising at least one compound of formula I as active ingredient. The pharmaceutical formulation may include one or more excipients and / or carrier materials. Furthermore, the formulation may contain any other active ingredient that inhibits the function of the choline kinase enzyme.

  These excipients, carrier substances and auxiliary substances are pharmaceutically, pharmacologically so that they can be combined with the other ingredients of the formulation or formulation and have no detrimental effect on the organism to be treated. It must be acceptable. These pharmaceutical compositions or formulations include those suitable for oral or parenteral administration (including subcutaneous, intradermal, intramuscular and intravenous administration), but the best route of administration depends on the patient's condition Different. These formulations may be in a single dose form. These formulations can be prepared according to methods known in the pharmaceutical field. The amount of active substance administered will vary according to the therapeutic details.

  The present invention also provides a process for the preparation of compounds of formula I. For this purpose of the invention, there are two different embodiments depending on whether the compounds of formula I have the same aminopyridinium group or different aminopyridinium groups.

a) A method for obtaining a compound of formula I having the same aminopyridinium group: This method comprises the corresponding heterocyclic derivative of formula VII and the dihalogenated derivative AX ₂ (where X is a halogen atom: Cl, Br or I In a 2: 1 molar amount in an organic solvent. This reaction is preferably carried out in butanone in a sealed tube at a temperature of 90-110 ° C.
b) A method for obtaining a compound of formula I in which the aminopyridinium group is different: this method comprises the corresponding heterocyclic derivative of formula VII and the dihalogenated derivative AX ₂ (where X is a halogen atom: Cl, Br or I Is represented in an organic solvent in a 1: 1 molar amount to give a monoquaternized product which is dissolved in the first organic solvent so that the preformed monoquaternized salt can be dissolved. Reacting again with another different heterocyclic derivative molecule in a 1: 1 molar ratio using another more polar organic solvent. The first step of this reaction is preferably performed at a temperature of 90-110 ° C. in butanone in a sealed test tube, and the second step is preferably performed at a temperature of 90-110 ° C. in ethanol in a sealed test tube. .

［式中、
Ｒ_１は、Ｈ、および場合によってトリフルオロメチル、ヒドロキシルまたはアルコキシルにより置換されていてもよいＣ_１−６アルキルからなる群から選択される基を表し；
Ｒ_２は、場合によってハロゲン、トリフルオロメチル、ヒドロキシル、Ｃ_１−６アルキル、アミノまたはアルコキシルにより置換されていてもよいアリール基を表し；
Ｒ_３は、Ｈ、ハロゲン、トリフルオロメチル、ヒドロキシル、アミノ、アルコキシル、および場合によってトリフルオロメチル、ヒドロキシル、アミノまたはアルコキシルにより置換されていてもよいＣ_１−６アルキルからなる群から選択される基か、あるいはＲ_４とともに、場合によってハロゲン、トリフルオロメチル、ヒドロキシル、Ｃ_１−６アルキル、アミノまたはアルコキシルにより置換されていてもよい−ＣＨ＝ＣＨ−ＣＨ＝ＣＨ−基のいずれかを表し；
Ｒ_４は、Ｈ、および場合によってハロゲン、トリフルオロメチル、ヒドロキシル、アミノまたはアルコキシルにより置換されていてもよいＣ_１−６アルキルからなる群から選択される基か、あるいはＲ_３とともに、場合によってハロゲン、トリフルオロメチル、ヒドロキシル、Ｃ_１−６アルキル、アミノまたはアルコキシルにより置換されていてもよい−ＣＨ＝ＣＨ−ＣＨ＝ＣＨ−基のいずれかを表す］
の化合物を提供する。 Finally, for its last purpose, the present invention relates to a compound of formula VII involved as starting compound in the process for the preparation of compounds of formula I:

[Where:
R ₁ represents a group selected from the group consisting of H and C _1-6 alkyl optionally substituted by trifluoromethyl, hydroxyl or alkoxyl;
R ₂ represents an aryl group optionally substituted by halogen, trifluoromethyl, hydroxyl, C _1-6 alkyl, amino or alkoxyl;
R ₃ is a group selected from the group consisting of H, halogen, trifluoromethyl, hydroxyl, amino, alkoxyl, and C _1-6 alkyl optionally substituted by trifluoromethyl, hydroxyl, amino or alkoxyl Or together with R ₄ represents any of the —CH═CH—CH═CH— groups optionally substituted by halogen, trifluoromethyl, hydroxyl, C _1-6 alkyl, amino or alkoxyl;
R ₄ is H and a group selected from the group consisting of C _1-6 alkyl optionally substituted by halogen, trifluoromethyl, hydroxyl, amino or alkoxyl, or together with R ₃ optionally halogen , Represents a —CH═CH—CH═CH— group optionally substituted by trifluoromethyl, hydroxyl, C _1-6 alkyl, amino or alkoxyl]
Of the compound.

の化合物である。
以下の実施例は本発明の例として示すものである。 Among the compounds of formula VII, preferably the compound is of formula VIII:

It is this compound.
The following examples are given as examples of the present invention.

Production example
Compound 1 (Code ACG560B): 1,1 ′-(Benzene-1,3-diylmethylene) bis [4- (4-chloro-N-methylanilino) pyridinium] dibromide in dry butanone (40 ml), 4- (4- A mixture of chloro-N-methylaniline) pyridine (125 mg, 0.57 mmol) and 1,3-bis (bromomethyl) benzene (75 mg, 0.28 mmol) was heated in a sealed test tube at 100 ° C. for 144 hours. After filtration and washing well with butanone, EtOAc and Et ₂ O, compound 1 was obtained as a pure white solid (125.2 mg, 62.7%); mp: 197-198 ° C. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 8.48 (d, 4H, H-2,6 _pyr , J = 6.6); 7.64 (d, 4H, H-3,5 _anil , J = 8.6); 7.57 (s, 1H, H-2ph); 7.45 (d, 5H, H-2,6 _anil and H-5ph; J = 8.6); 7.37 (d, 2H, H-4,6 _ph , J = 7.7); 6.95 (bs, 4H, H-3,5 _pyr ); 5.49 (s, 4H, CH ₂ N ⁺ ); 3.46 (s, 6H, Me). ¹³ C-NMR (100 MHz, DMSO-d ₆ ) δ 156.20 (C-4 _pyr ); 142.75 (C-2,6 _pyr ); 141.96 (C-1 _anil ); 136.18 (C- 1.3 _ph ); 132.78 (C-4 _anil ); 130.50 (C-3,5 _anil ) 129.73 (C-5 _ph ); 128.37 (C-2,6 _anil ); 128.18 (C-4,6 _ph ); 127.89 (C-2 _ph ); 109.15 (C-3,5 _pyr ); 59.16 (CH ₂ N ⁺ ); 41.42 (Me). HRMS (m / z): C ₃₂ H ₃₀ N ₄ Calculated as Cl ₂ Br (M − Br) ⁺ : 619.1031; Found: 619.1031. C ₃₂ H ₃₀ N ₄ Analysis as Cl ₂ Br ₂ · 1H ₂ O .: Calculated: C 53.43; H 4.56; N 7.63%. Found: C 53.14; H 4.48; N 7.79%.

Compound 2 (Code ACG416B): 1,1 ′-(biphenyl-3,3′-diylmethylene) bis [4- (N-methylanilino) pyridinium] dibromide in dry butanone (40 ml), 4- (N-methylaniline) A mixture of pyridine (216 mg, 1.17 mmol) and 3,3′-bis (bromomethyl) biphenyl (200 mg, 0.58 mmol) was heated in a sealed tube at 100 ° C. for 24 hours. After filtration and washing well with butanone, the solid product was purified by recrystallization from MeOH and the residue was triturated with Et ₂ O. Compound 2 was obtained as a white solid (294 mg, 71.5%); mp: 124-125 ° C. ¹ H-NMR (300 MHz, CD ₃ OD) δ 8.35 (bs, 4H, H-2,6 _pyr ); 7.84 (s, 2H, H-2 _ph ); 7.67 (d, 2H, H-6 _ph , J = 7.7); 7.56 (t, 4H, A-3,5 _anil , J = 7.6); 7.50-7.44 (m, 4H, H-5 _ph and H-4 _anil ); 7.39 (d, 2H, H- 4 _ph , J = 7.7); 7.33 (d, 4H, H-2,6 _anil , J = 7.5); 6.95 (bs, 4H, H-3,5 _pyr ); 5.47 (s, 4H, CH ₂ N ⁺ ); 3.51 (s, 6H, Me) 13 C-NMR (75 MHz, CD 3 OD) δ 158.48 (C-4 pyr);. 144.82 (C-1 anil); 143.80 (C-2.6 pyr); 142.60 ( C.1 _ph ); 136.82 (C-3 _ph ); 132.01 (C-3,5 _anil ); 131.14 (C-5 _ph ); 130.12 (C-4 _anil ); 128.99 (C-4 _ph ); 128.82 ( C-6 _ph ); 128.58 (C-2 _ph ); 127.52 (C-2,6 _anil ); 110.29 (C- 3,5 _pyr ); 61.97 (CH ₂ N ⁺ ); 41.42 (Me) .HRMS (m / z): Calculated as C ₃₈ H ₃₆ N ₄ Br (M − Br) ⁺ : 627.2123; Found: 627.2122. Analysis as C ₃₈ H ₃₆ N ₄ Br ₂ · 2.5H ₂ O .: Calculated: C 60.56; H 5.48; N 7.43%. Found: C 60.70; H 5.83; N 7.20%.

Compound 3 (Code ACG548B): 1,1 ′-(biphenyl-3,3′-diylmethylene) bis [4- (4-chloro-N-methylanilino) pyridinium] dibromide in dry butanone (40 ml) 4- (4 A mixture of -chloro-N-methylaniline) pyridine (235 mg, 1.07 mmol) and 3,3′-bis (bromomethyl) biphenyl (183 mg, 0.53 mmol) was heated in a sealed test tube at 100 ° C. for 24 hours. After filtration and washing well with CHCl ₃ , the solid product was purified by recrystallization from MeOH after adding Et ₂ O until cloudy. Compound 3 was obtained as a white solid (205 mg, 49.7%); mp: 279-280 ° C. ¹ H-NMR (300 MHz, DMSO-d ₆ ) δ 8.57 (d, 4H, H-2,6 _pyr , J = 6.5); 7.88 (s, 2H, H- 2 _ph ); 7.67 (d, 2H, H-6 _ph , J = 7.7); 7.61 (d, 4H, H-3,5 _anil , J = 8.6); 7.51 (t, 2H, H-5 _ph , J = 7.7); 7.42 (d, 6H, H-4 _ph and H- 2,6 _anil , J = 8.6); 6.99 (bs, 4H, H-3,5 _pyr ); 5.51 (s, 4H, CH ₂ N ⁺ ); 3.43 (s, 6H, Me ¹³ C-NMR (75 MHz, DMSO-d ₆ ) δ 156.20 (C-4 _pyr ); 142.72 (C-2,6 _pyr ); 142.05 (C-1 _anil ); 140.01 (C-1 _ph ); 136.20 (C-3 _ph ); 132.79 (C-4 _anil ); 130.53 (C-3,5 _anil ); 129.73 (C-5 _ph ); 128.47 (C-2,6 _anil ); 127.51 (C-4 _ph ); 127.14 (C-6 _ph ); 127.04 (C-2 _ph ); 109.20 (C- 3,5 _pyr ); 59.55 (CH ₂ N ⁺ ); 40.73 (Me) .HRMS (m / z): C ₃₈ Calculated as H ₃₄ N ₄ Cl ₂ Br (M − Br) ⁺ : 695.1344; Found: 695.1344. Analysis as C ₃₈ H ₃₄ N ₄ Cl ₂ Br ₂ • 1.2H ₂ O: Calculated: C 57.12; H 4.59; N 7.01%. Found: C 57.55; H 4.99; N 6.97%.

Compound 4 (Code ACG604A): 1,1 ′-(biphenyl-3,3′-diylmethylene) bis [4- (3,5-dichloro-N-methylanilino) pyridinium] dibromide in dry butanone (40 ml), 4- A mixture of (3,5-dichloro-N-methylaniline) pyridine (200 mg, 0.80 mmol) and 3,3′-bis (bromomethyl) biphenyl (136 mg, 0.40 mmol) was added in a sealed test tube at 100 ° C. at 72 ° C. Heated for hours. After filtration and washing well with butanone Et ₂ O, compound 4 was obtained as a pure white solid (270 mg, 79.7%); mp: 312-313 ° C. ¹ H-NMR (300 MHz, DMSO-d ₆ ) δ 8.63 (d, 4H, H-2,6 _pyr , J = 7.1); 7.92 (s, 2H, H-2 _ph ); 7.75 (s, 2H, H-4 _anil ); 7.70 (d, 2H, H-6 _ph , J = 7.6); 7.62 (d, 4H, H-2,6 _anil , J = 1.8); 7.53 (t, 2H, H-5 _ph , J = 7.6); 7.45 (d, 2H, H-4 _ph , J = 7.6); 7.04 (d, 4H, H-3,5 _pyr , J = 7.1); 5.56 (s, 4H, CH ₂ N ⁺ ); 3.44 (s, 6H, Me) 13 C-NMR (75 MHz, DMSO-d 6) δ 156.20 (C-4 pyr);. 145.27 (C-1 anil); 142.86 (C-2,6 pyr) 140.08 (C-1 _ph ); 136.11 (C-3 _ph ); 135.34 (C-3,5 _anil ); 129.70 (C-5 _ph ); 128.33 (C-4 _anil ); 127.55 (C-4 _ph ) ; 127.14 (C-6 _ph ); 127.07 (C-2 _ph ); 125.97 (C-2,6 _anil ); 109.53 (C- 3,5 _pyr ); 59.65 (CH ₂ N ⁺ ); 40.59 (Me). HRMS (m / z): Calculated as C ₃₈ H ₃₂ N ₄ Cl ₄ Br (M-Br) ⁺ : 763.0564; Found: 763.0563. As C ₃₈ H ₃₂ N ₄ Cl ₄ Br ₂ · 0.1H ₂ O Analysis: Calculated: C 53.81; H 3.81; N 6.60%. Found: C 53.41; H 4.19; N 6.25%.

Compound 5 (Code RSM964A): 1,1 ′-(biphenyl-3,3′-diylmethylene) bis [4- (4-chloro-N-methylanilino) quinolinium] dibromide in dry butanone (40 ml) 4- (4 A mixture of -chloro-N-methylanilino) quinoline (212 mg, 0.78 mmol) and 3,3′-bis (bromomethyl) benzene (134 mg, 0.39 mmol) was heated in a sealed test tube at 100 ° C. for 72 hours. After filtration and washing well with butanone, EtOAc and Et ₂ O, compound 5 was obtained as a pure yellowish solid (134 mg, 40%); mp: 217-218 ° C. ¹ H-NMR (300 MHz, DMSO-d ₆ ): δ 9.24 (d, J = 7.4, 2H, H-2 _quin ); 8.18 (d, J = 8.9, 2H, H-8 _quin ); 7.84 (s , 2H, H-2 _ph ); 7.63 (d, J = 7.5, 2H, H-5 _quin ); 7.56-7.43 (m, 18H, H-5,6 _ph , H-2,3,5,6 _anil , H- _{3,6,7 quin} ); 7.23 (d, J = 7.4, 2H, H-4 _ph ); 6.08 (s, 4H, N ⁺ -CH ₂ ); 3.74 (s, 6H, Me). ¹³ C-NMR (100 MHz, DMSO-d ₆ ): δ 157.87 (C-4 _quin ); 147.46 (C-2 _quin ); 146.42 (C-1 _anil ); 140.03 (C-1 _ph ); 138.83 (C- 8a _quin ); 135.61 (C-3 _ph ); 133.50 (C-7 _quin ); 131.69 (C-4 _anil ); 130.27 (C-3,5 _anil ); 129.62 (C-5 _ph ); 127.35 (C- _{6 ph); 127.18 (C-} 2,6 anil); 126.73 (C-6 quin); 126.09 (C- 4 ph); 125.87 (C-5 quin); 125.67 (C-2 ph); 119.65 (C- 4a _quin ); 119.14 (C-8 _quin ); 107.10 (C-3 _quin ); 57.28 (N + -CH ₂ ); 44.94 (Me) .HRMS (m / z): C ₄₆ H ₃₈ N ₄ Cl ₂ Br ₂ Calculated as [(M-Br)] ⁺ : 795.1657. Found: 795.1656. Analysis as C ₄₆ H ₃₈ N ₄ Cl ₂ Br ₂ · 3H ₂ O: Calculated: C 59.31; H 4.76; N 6.01% Actual value: C 59.24
; H 4.70; N 5.65%.

Compound 6 (Code RSM820C): 1,1 ′-(biphenyl-3,3′-diylmethylene) bis [4- (4-chloro-N-methylanilino) -7-chloroquinolinium] dibromide dried butanone (40 ml) A mixture of 7-chloro-4- (4-chloro-N-methylanilino) quinoline (300 mg, 0.98 mmol) and 3,3′-bis (bromomethyl) biphenyl (168 mg, 0.49 mmol) in a sealed test tube. And heated at 100 ° C. for 72 hours. After filtration and thorough washing with butanone and CHCl ₃ , the solid product was purified by recrystallization from EtOH or EtOH / MeOH after adding Et ₂ O until cloudy. Compound 6 was obtained as a yellowish solid (154 mg, 45%); mp: 220-221 ° C.
¹ H-NMR (300 MHz, DMSO-d ₆ ): δ 9.19 (d, J = 7.5, 2H, H-2 _quin ); 8.29 (d, J = 1.7, 2H, H-8 _quin ); 7.85 (s , 2H, H-2 _ph ); 7.64 (d, J = 7.2, 2H, H-5 _quin ); 7.57-7.45 (m, 16H, H-5,6 _ph , H-2,3,5,6 _anil , H-3,6 _quin ); 7.25 (d, J = 7.7, 2H; H-4 _ph ); 6.08 (s, 4H, N + -CH ₂ ); 3.73 (s, 6H, Me). ¹³ C-NMR (75 MHz, DMSO-d ₆ ): δ 157.68 (C-4 _quin ); 148.01 (C-2 _quin ); 146.14 (C-1 _anil ); 140.14 (C-1 _ph ); 139.85 (C-8a _quin ) 138.48 (C-7 _quin ); 135.51 (C-3 _ph ); 132.11 (C-4 _anil ); 130.50 (C-3,5 _anil ); 129.80 (C-5 _ph ); 129.45. (C-6 _ph ); 127.32 (C-2,6 _anil ); 126.89 (C-6 _quin ); 126.12 (C-4 _ph ); 125.91 (C-5 _quin ); 125.82 (C-2 _ph ); 118.48 (C-8 _quin) ); 118.35 (C-4a _quin ); 107.38 (C-3 _quin ); 57.14 (N ⁺ -CH ₂ ); 45.18 (Me) .HRMS (m / z): C ₄₆ H ₃₆ N ₄ Cl ₄ Br ₂ [ .. (M - HBr - Br )] + as calculated: 783.1616 _{Found: 783.1616 C 46 H 36 N 4} Cl 4 Br 2 · 1.5H analysis as ₂ O: calculated: C 56.76; H 4.04; N 5.76%. Found: C 56.72; H 4.18; N 5.71%.

Compound 7 (Code RSM 932A): 1,1 ′-(biphenyl-4,4′-diylmethylene) bis [4- (4-chloro-N-methylanilino) quinolinium] dibromide in dry butanone (40 ml) 4- (4 A mixture of -chloro-N-methylaniline) quinoline (240 mg, 0.89 mmol) and 4,4′-bis (bromomethyl) biphenyl (152 mg, 0.44 mmol) was heated in a sealed test tube at 100 ° C. for 72 hours. After filtration and washing well with butanone, compound 7 was obtained as a pure yellowish solid (121 mg, 30%); mp: 255-257 ° C. ¹ H-NMR (300 MHz, DMSO-d ₆ ): δ 9.19 (d, J = 7.4, 2H, H-2 _quin ); 8.12 (d, J = 8.9, 2H, H-8 _quin ); 7.83 (pst , J = 7.5, 2H, H-7 _quin ); 7.66 (d, J = 8.2, 2H, H-5 _quin ); 7.55 (d, J = 8.8, 4H, H-3,5 _anil ); 7.44 (d , J = 8.9, 4H, H-2,6 _anil ); 7.56-7.39 (m, 12H, H-2,3,5,6 _ph , H-3 _quin , H-6 _quin ); 6.05 (s, 4H _{^{, N + -CH 2); 3.73}} (s, 6H, Me) 13 C-NMR (75 MHz, DMSO-d 6):. δ 157.86 (C-4 quin); 147.41 (C-2 quin); 146.40 ( C-1 _anil ); 139.11 (C-1 _ph ); 138.78 (C-8a _quin ); 134.30 (C-4 _ph ); 133.47 (C-7 _quin ); 131.69 (C-4 _anil ); 130.26 (C- 3,5 _anil ); 127.34 (C-3,5 _ph ); 127.18 (C-2,6 _anil ), (C-2,6 _ph ); 127.08 (C-6 _quin ); 126.08 (C-5 _quin ) 119.65 (C-4a _quin ); 119.12 (C-8 _quin ); 107.06 (C-3 _quin ); 56.94 (N ⁺ -CH ₂ ); 44.94 (Me) _.HRMS (m / z): C ₄₆ H ₃₈ Calculated as N ₄ Cl ₂ Br ₂ [(M-Br)] ⁺ : 795.1657. Found: 795.1658. C ₄₆ H ₃₈ Analysis as N ₄ Cl ₂ Br ₂ · 2H ₂ O: Calculated: C 60.48; H 4.63; N 6.13%. Found: C 60.0
6; H 4.48; N 5.87%.

Compound 8 (Code RSM824B): 1,1 ′-(biphenyl-4,4′-diylmethylene) bis [4- (4-chloro-N-methylanilino) -7-chloroquinolinium] dibromide dried butanone (100 ml) A mixture of 7-chloro-4- (4-chloro-N-methylanilino) quinoline (300 mg, 0.98 mmol) and 4,4′-bis (bromomethyl) biphenyl (168 mg, 0.49 mmol) in a sealed test tube. And heated at 100 ° C. for 72 hours. After filtration and washing well with butanone, compound 8 was obtained as a pure yellowish solid (195 mg, 48%); mp: 276-277 ° C. ¹ H-NMR (400 MHz, DMSO-d ₆ ): δ 9.14 (d, J = 7.4, 2H, H- 2 _quin ); 8.23 (d, J = 1.6, 2H, H-8 _quin ); 7.73 (d , J = 8.3, 2H, H-5 _quin ); 7.69 (d, J = 8.4, 4H, H-2,6 _ph ); 7.56 (d, J = 8.8, 4H, H- 3,5 _anil ); 7.46 (d, J = 8.9, 4H, H-2,6 _anil ); 7.50-7.46 (m, 6H, H-6 _quin , H-3 _quin ); 7.41 (d, J = 8.4, 4H, H-3, _{. 5 ph); 6.04 (s} , 4H, N + - CH 2); 3.73 (s, 6H, Me) 13 C-NMR (100 MHz, DMSO-d 6): δ 157.69 (C- 4 quin); 147.98 (C-2 _quin ); 146.13 (C-1 _anil ); 139.82 (C-8a _quin ); 139.21 (C-1 _ph ); 138.51 (C-7 _quin ); 134.22 (C-4 _ph ); 132.14 (C -4 _anil ); 130.50 (C-3,5 _anil ); 129.45 (C-2,6 _anil ); 127.54 (C-3,5 _ph ); 127.33 (C-6 _quin ); 127.23 (C-2,6 _ph ); 126.52 (C-5 _quin ); 118.47 (C-8 _quin ); 118.35 (C-4 a _quin ); 107.33 (C-3 _quin ); 56.83 (N ⁺ -CH ₂ ); 45.19 (Me): HRMS (m / e): Calculated as C ₄₆ H ₃₆ N ₄ Cl ₄ Br ₂ [(M-HBr-Br)] ⁺ : 783.1616. Found: 783.1614. As C ₄₆ H ₃₆ N ₄ Cl ₄ Br ₂ Analysis: Calculated: C 58.3
8; H 3.83; N 5.92%. Found: C 58.73; H 3.96; N 5.74%.

Compound 9 (code RSM936A): 1,1 ′-[ethylenebis (benzene-1,4-diylmethylene)] bis [4- (4-chloro-N-methylanilino) quinolinium] dibromide in dry butanone (40 ml) 4 A mixture of-(4-chloro-N-methylanilino) quinoline (204 mg, 0.76 mmol) and 4,4'-bis (bromomethyl) bibenzyl (140 mg, 0.37 mmol) was heated in a sealed test tube at 100 ° C for 72 hours. did. After filtration and washing well with butanone and CHCl ₃ , compound 9 was obtained as a pure yellowish solid (70 mg, 20%); mp: 212-214 ° C. ¹ H-NMR (300 MHz, DMSO-d ₆ ): δ 9.19 (d, J = 7.4, 2H, H-2 _quin ); 8.10 (d, J = 8.9, 2H, H-8 _quin ); 7.82 (pst , J = 7,5, 2H, H-7 _quin ); 7.54 (d, J = 8.8, 4H, H-3,5 _anil ); 7.44 (d, J = 8.9, 4H, H-2,6 _anil ) 7.52-7.39 (m, 6H, H-3 _quin , H-5 _quin , H-6 _quin ); 7.24 (s, 8H, H-2,3,5,6 _ph ); 5.98 (s, 4H, N _{^{+ -CH 2); 3.73 (s}} , 6H, Me); 2.80 (s, 4H, CH 2 -Ph) 13 C-NMR (100 MHz, DMSO-d 6):. δ 157.80 (C-4 quin); 147.34 (C-2 _quin ); 146.44 (C-1 _anil ); 141.55 (C-1 _ph ); 138.74 (C-8a _quin ); 133.3.6 (C- _quin 7); 132.32 (C.4 _ph ); 131.63 (C-4 _anil ); 130.25 (C-3,5 _anil ); 128.79 (C-3,5 _ph ); 127.26 (C-6 _quin ); 127.17 (C-2,6 _anil ); 126.74 (C- 2,6 _ph ); 126.04 (C-5 _quin ); 119.66 (C-4 a _quin ); 119.19 (C-8 _quin ); 107.06 (C-3 _quin ); 57.10 (N ⁺ -CH ₂ ); 44.93 (Me ); 36.22 (CH ₂ -Ph). HRMS (m / z): Calculated as C ₄₈ H ₄₂ N ₄ Cl ₂ Br ₂ [(M-Br)] ⁺ : 823.1970. Found: 823.1970. C ₄₈ H Analysis as ₄₂ N ₄ Cl ₂ Br ₂ · 1H ₂ O .: Calculated: C 62.42; H 4.80; N 6.07%. Found: C 62.2 9; H 4.59; N 6.09%.

Compound 10 (Code RSM828B): 1,1 ′-[ethylenebis (benzene-1,4-diylmethylene)] bis [4- (4-chloro-N-methylanilino) -7-chloroquinolinium] dibromide dried butanone (40 ml) was sealed with a mixture of 7-chloro-4- (4-chloro-N-methylanilino) quinoline (300 mg, 0.98 mmol) and 4,4′-bis (bromomethyl) bibenzyl (182 mg, 0.49 mmol). Heated in a test tube at 100 ° C. for 72 hours. After filtration and washing well with butanone, compound 10 was obtained as a pure yellowish solid (229 mg, 48%); mp: 256-257 ° C. ¹ H-NMR (400 MHz, DMSO-d ₆ ): δ 9.11 (d, J = 7.4, 2H, H- 2 _quin ); 8.18 (d, J = 1.5, 2H, H-8 _quin ); 7.55 (d , J = 8.8, 4H, H- 3,5 _anil ); 7.46 (d, J = 8.8, 4H, H-2,6 _anil ); 7.56-7.44 (m, 6H, H-3 _quin , H-5 _quin , H-6 _quin ); 7.24 (s, 8H, H-2,3,5,6 _ph ); 5.97 (s, 4H, N ⁺ -CH ₂ ); 3.72 (s, 6H, Me); 2.82 (s ^{_{, 4H, CH 2 -Ph) 13}} C-NMR (100 MHz, DMSO- d 6):. δ 157.63 (C-4 quin); 147.91 (C-2 quin); 146.16 (C-1 anil); 141.74, 139.75 and 138.88 (C-7 _quin , C-8a _quin and C-4 _ph ); 132.20 (C-4 _anil ); 132.08 (C-1 _ph ); 130.50 (C-3,5 _anil ); 129.39 (C- 6 _quin ); 128.99 (C- 3,5 _ph ); 127.32 (C-2,6 _anil ); 126.90 (C-2,6 _ph ); 126.48 (C-5 _quin ); 118.55 (C-8 _quin ); 118.35 (C-4a _quin ); 107.32 (C-3 _quin ); 57.02 (N ⁺ -CH ₂ ); 45.17 (Me); 36.33 (CH ₂ -Ph) .HRMS (m / z): C ₄₈ H ₄₀ N Calculated as ₄ Cl ₄ Br ₂ [(M-HBr-Br)] ⁺ : 811.1927. Found: 811.1926. Analysis as C ₄₈ H ₄₀ N ₄ Cl ₄ Br ₂ · 2H ₂ O: Calculated: C 57.05 ; H 4.39; N 5
.54%. Found: C 57.14; H 4.07; N 5.46%.

以下の出発材料は個々の参照文献に記載されている方法によって作製した。 Reagent Preparation The compound α, α′-dibromo-m-xylene is a commercial product provided by Sigma-Aldrich Quimica SA (Avenida Valdelaparra No. 51-53, 28100 Alcobendas (Madrid)).

The following starting materials were made by the methods described in the individual references.

Werner, W. J. Org. Chem. 17, 523-528 (1952)
２．− ４，４’−ビス（ブロモメチル）ビフェニル

Szendey, G. L., Munnes, S. Chem. Ber. 94, 38-42 (1961); Staab, H. A., Haenel, M. Chem. Ber. 106, 2190-2202 (1973)
３．− ビス−ｐ−（ブロモメチル）ビベンジル

Cram, D. J., Steinberg, J. J. Am. Chem. Soc. 73, 5691-5704 (1951)
４．− ４−（Ｎ−メチルアニリノ）ピリジン

Campos, J., Nunez, M. C., Sanchez, R., Gomez-Vidal, J. A., Rodriguez-Gonzalez, A., Banez, M., Gallo, M. A., Lacal, J. C., Espinosa, A. Bioorg. & Med. Chem. 10, 2215-2231 (2002)
５．− ４−（４−クロロ−Ｎ−メチルアニリノ）ピリジン

Conejo-Garcia, A., Campos, J., Sanchez, R., Rodriguez-Gonzalez, A., Lacal, J. C., Gallo, M. A., Espinosa, A. Eur. J. Med. Chem. 38, 109-116 (2003)
６．−４−（３，５−ジクロロ−Ｎ−メチルアニリノ）ピリジン

1. -3,3'-bis (bromomethyl) biphenyl

Werner, WJ Org. Chem. 17, 523-528 (1952)
2. -4,4'-bis (bromomethyl) biphenyl

Szendey, GL, Munnes, S. Chem. Ber. 94, 38-42 (1961); Staab, HA, Haenel, M. Chem. Ber. 106, 2190-2202 (1973)
3. -Bis-p- (bromomethyl) bibenzyl

Cram, DJ, Steinberg, JJ Am. Chem. Soc. 73, 5691-5704 (1951)
4). 4- (N-methylanilino) pyridine

Campos, J., Nunez, MC, Sanchez, R., Gomez-Vidal, JA, Rodriguez-Gonzalez, A., Banez, M., Gallo, MA, Lacal, JC, Espinosa, A. Bioorg. & Med. Chem . 10, 2215-2231 (2002)
5. 4- (4-Chloro-N-methylanilino) pyridine

Conejo-Garcia, A., Campos, J., Sanchez, R., Rodriguez-Gonzalez, A., Lacal, JC, Gallo, MA, Espinosa, A. Eur. J. Med. Chem. 38, 109-116 ( 2003)
6). -4- (3,5-dichloro-N-methylanilino) pyridine

  This compound has so far been synthesized by Conejo-Garcia, A., Campos, J., Sanchez, R., Rodriguez-Gonzalez, A., Lacal, JC, Gallo, MA, Espinosa, A. Eur. J. Med. Chem. Prepared from 4-chloropyridine hydrochloride and 4- (3,5-dichloro-N-methylanilino) pyridine according to the method described in 38, 109-116 (2003). On the other hand, 3,5-dichloro-N-methylanilino has been studied in the following studies: Leeson, PD, Baker, R., Carling, RW, Curtis, NR, Moore, KW, Williams, BJ, Foster, AC, Donald, AE, Kemp , JA, Marshall, GRJ Med. Chem. 34, 1243-1252 (1991).

参照：Chi, AND. F.; Chu, T. I. Record (Peking), 1, 45 (1957); Chem. Abstract, 52, 6321 a,b (1957)
８．− ジエチル−４，４）’−ビス（ブロモメチル）−［２，２’］ビチアゾリル−５，５’−二カルボン酸塩

参照：Lehn, J.-M.; Regnouf de Vains, J.-B. Tetrahedron Lett., 30, 2209-2212 (1989)
９．− ６，６’−ビス（ブロモメチル）−［２，２’］ビピリジン

参照：Rodriguez-Ubis, J.-C.; Alpha, B.; Plancherel, D.; Lehn, J.-M. Helv. Chim. Acta, 67, 2264 (1984)
１０．− ６，６’−ビス（ブロモメチル）−４，４’−ジメチル−［２，２’］ビピリミジニル

参照：Lehn, J.-M.; Regnouf de Vains, J.-B. Tetrahedron Lett., 30, 2209-2212 (1989) 7). -4,4'-bis (chloromethyl)-[2,2 '] bithiazolyl

Reference: Chi, AND. F .; Chu, TI Record (Peking), 1, 45 (1957); Chem. Abstract, 52, 6321 a, b (1957)
8). -Diethyl-4,4) '-bis (bromomethyl)-[2,2'] bithiazolyl-5,5'-dicarboxylate

Reference: Lehn, J.-M .; Regnouf de Vains, J.-B. Tetrahedron Lett., 30, 2209-2212 (1989)
9. 6,6′-bis (bromomethyl)-[2,2 ′] bipyridine

Reference: Rodriguez-Ubis, J.-C .; Alpha, B .; Plancherel, D .; Lehn, J.-M. Helv. Chim. Acta, 67, 2264 (1984)
10. 6,6′-bis (bromomethyl) -4,4′-dimethyl- [2,2 ′] bipyrimidinyl

Reference: Lehn, J.-M .; Regnouf de Vains, J.-B. Tetrahedron Lett., 30, 2209-2212 (1989)

の化合物は、氷酢酸中、還流下で、４−アニリンまたはキノリン誘導体を対応する４−クロロ−アニリンと反応させることで作製できる。冷却後、この溶液を水酸化ナトリウム溶液で塩基性とし、その後、得られた懸濁液を濃縮し、フラッシュクロマトグラフィーにより精製する。 Formula VII for creating new starting materials :

Can be prepared by reacting 4-aniline or a quinoline derivative with the corresponding 4-chloro-aniline in glacial acetic acid under reflux. After cooling, the solution is made basic with sodium hydroxide solution, after which the resulting suspension is concentrated and purified by flash chromatography.

Formula VIII:

の化合物を得る例を以下に示す。

An example of obtaining the compound is shown below.

Compound VIIIA
4- (4-Chloro-N-methylanilino) quinoline A solution of 4-chloroquinoline (5 mmol) and 4-chloro-N-methylaniline (10 mmol) in glacial acetic acid (15 ml) was heated to reflux under a stream of argon for 3 hours. did. After cooling, the solution is basified with 10% NaOH solution to pH = 10 and the resulting suspension is concentrated on a rotary evaporator and purified by flash chromatography (9: 1, CH ₂ Cl ₂ : MeOH). The target molecule was obtained as a yellowish syrup (97%). ¹ H-NMR (400MHz, CDCl3): δ 8.10 (d, J = 8.5, 1H, H-2 _quin ); 7.70 (d, J = 8.5, 1H, H-5 _quin ); 7.65 (t, J = 7.9 , 1H, H- 7 _quin ); 7.38 (t, J = 8.5, 1H, H-6 _quin ); 7.35 (d, J = 7.9, 1H, H- 8 _quin ); 7.17 (d, J = 8.9, 2H , H-3,5 _anil ); 7.14 (d, J = 8.5, 1H, H-3 _quin ); 6.76 (d, J = 8.9, 2H, H-2,6 _anil ); 3.45 (s, 6H, Me ¹³ C-NMR (100 MHz, CDCl3): δ 153.37 (C-4 _quin ); 151.16 (C-2 _quin ); 150.01 (C- 1 _anil ); 148.17 (C-8a _quin ); 135.02 (C- 4 _anil ); 130.07 (C-7 _quin ); 129.52 (C-6 _quin ); 129.29 (C-3,5 _anil ); 126.26 (C-4a _quin ); 126.07 (C-5 _quin ); 124.40 (C- 8 _quin ); 119.79 (C-2,6 _anil ); 115.08 (C-3 _quin ); 41.75 (Me). HRMS (m / z): C ₁₆ H ₁₃ N ₂ Cl [(M + H)] ⁺ Calculated value: 269.0845. Found: 269.0845. Analysis as C ₁₆ H ₁₃ N ₂ Cl: Calculated: C 71.51; H 4.88; N 10.42%. Found: C 71.60; H 4.71; N 10.33%.

Compound VIIIB
7-Chloro-4- (4-chloro-N-methylanilino) quinoline A solution of 4,7-dichloroquinoline (5 mmol) and 4-chloro-N-methylaniline (10 mmol) in glacial acetic acid (15 ml) under a stream of argon. And heated at reflux for 3 hours. After cooling, the solution is basified with 10% NaOH solution to pH = 10 and the resulting suspension is concentrated on a rotary evaporator and purified by flash chromatography (9: 1, CH ₂ Cl ₂ : MeOH). Intermediate II was obtained as a yellowish syrup (59%).
¹ H-NMR (300 MHz, CH3OD): δ 8.66 (d, J = 7.1, 1H, H-2 _quin ); 7.94 (d, J = 2.0, 1H, H-8 _quin ); 7.53 (d, J = 8.8, 2H, H- 3,5 _anil ); 7.41-7.37 (m, 2H, H-5,6 _quin ); 7.47 (d, J = 8.8, 2H, H- 2,6 _anil ); 7.32 (d, . J = 7.1, 2H, H -3 quin); 3.76 (s, 3H, Me) 13 C-NMR (75 MHz, CH3OD): δ 159.86 (C-4 quin); 147.63 (C-7 quin); 143.86 (C- 2 _quin ); 141.46 (C-1 _anil ); 140.56 (C-8a _quin ); 135.02 (C-4 _anil ); 132.01 (C-3,5 _anil ); 129.92 (C-6 _quin ); 128.58 (C-2,6 _anil ); 127.98 (C-5 _quin ); 120.56 (C-8 _quin ); 118.71 (C-4a _quin ); 107.38 (C-3 _quin ); 45.74 (Me) .HRMS (m / z): Calculated as C ₁₆ H ₁₂ N ₂ Cl ₂ [(M + H)] ⁺ : 303.0456. Found: 303.0456. Analysis as C ₁₆ H ₁₂ N ₂ Cl ₂ .: Calculated: C 63.38; H 3.99; N 9.24%. Found: C 63.46; H 3.71; N 9.17%.

Ex vivo assay for human ChoK activity E. coli in buffer assay (100 mM Tris-HCl pH 8.0, 100 mM MgCl ₂ , 10 mM ATP and 200 μM choline in the presence of methyl [ ¹⁴ C] -choline chloride (50-60 μCi / mmol)) Recombinant choline kinase expressed in (E. coli) was used for ex vivo assays. The reaction was carried out at 37 ° C. for 30 minutes and stopped by adding ice-cooled trichloroacetic acid to a final concentration of 16%. These samples were washed with water saturated diethyl ether and lyophilized. These hydrophilic choline derivatives are separated on a thin layer chromatography plate according to the described method [Ramirez, A., Penalva, V., Lucas, L., Lacal, JC Oncogene 21, 937-946 (2002)]. did.

  These assays were performed using compounds 1-10 of the present invention, as well as compounds EC1-EC6 known in the state of the art, in particular ES 2 117 950. These results are summarized in Table II.

Cell Proliferation Assay HT-29 cells were seeded in 24-well plates (35 × 10 ³ cells / well) and incubated for 24 hours. These cells were then treated with various concentrations of ChoK inhibitor in normal culture medium. After 3 days, the wells were aspirated and both fresh media and an additional amount of drug were added and the cells were maintained for an additional 3 days. The crystal violet method [Gillies, RJ, Didier, N., Denton, M. Anal. Biochem. 159, 109-113 (1986)] was slightly modified [Hernandez-Alcoceba, R., Saniger, L., Campos, J., Nunez, MC, Khaless, F., Gallo, MA, Espinosa, A., Lacal, JC Oncogene, 15, 2289-2301 (1997)], the remaining cells in each well were quantified. . Briefly, cells were washed with TD buffer and fixed with 1% glutaraldehyde for 15 minutes. After washing again with TD, the cell nuclei were stained with 0.1% crystal violet for at least 30 minutes and washed 3 times with distilled water. The absorbed dye was resuspended in 10% acetic acid and the absorbance at 595 nm was measured with a spectrophotometer. The results obtained are summarized in the form of IC ₅₀ values, ie the concentration of compound required to produce 50% inhibition. This value is determined by iterative curve adjustment. Two values were determined for each point on the curve and the experiment was repeated two or three times to estimate the average value. When there was a difference of more than 50% between the two values, a third experiment was performed to determine the true value. IC ₅₀ values were used as potency measures to correlate the biological activity of the compounds with their chemical structure.

  These assays were performed using compounds 1-10 of the present invention, as well as compounds EC1-EC6 known in the state of the art, in particular ES 2 117 950. These results are summarized in Table II.

Toxicity assay Toxicity assays were performed using 1 month old Balb C mice weighing approximately 25-30 grams at the start of the experiment. Mice were inoculated with varying amounts of each compound from 0.1 mg / kg to 25 mg / kg daily for 5 consecutive days. After 5 doses, the mice were rested for 9 days and examined for both survival and general health, with particular attention to their hair, behavior, dietary habits and body weight. The 50% lethal dose was recorded as the corresponding toxicity IC ₅₀ . The results obtained with the new compounds show a clear increase in activity, in which case toxicity is reduced, as measured by their corresponding IC ₅₀ .

These assays were performed using compounds 1-10 of the present invention, as well as compounds EC1-EC6 known in the state of the art, in particular ES 2 117 950. These results are summarized in Table II.
Table II below summarizes the results obtained with the assays performed.

表IIのデータから、本発明の化合物は特許ＥＳ ２ １１７ ９５０の化合物よりも著しく低い毒性を有する一方で、培養腫瘍由来の細胞に対する抗増殖活性および免疫抑制マウスに接種したヒト腫瘍に対するin vivo抗腫瘍活性に同等以上の値を維持することが見て取れる。

From the data in Table II, the compounds of the present invention have significantly lower toxicity than the compounds of patent ES 2 117 950, while anti-proliferative activity against cells derived from cultured tumors and in vivo anti-human tumors inoculated into immunosuppressed mice. It can be seen that the tumor activity remains at or above the same value.

Claims (7)

Formula I:

[Where:
Q ⁻ represents a conjugated base of a pharmaceutically suitable organic or inorganic acid;
R ₁ and R ′ ₁ represent a methyl group;
R ₂ and R ′ ₂ independently of one another represent a phenyl group substituted by one or more halogen substituents ;
R ₃ and R ′ ₃ are, independently of each other, H, halogen, trifluoromethyl, hydroxyl, amino, alkoxyl, and optionally C _1-6 alkyl optionally substituted by trifluoromethyl, hydroxyl, amino or alkoxyl A group selected from the group consisting of or independently of each other, together with R ₄ and R ′ ₄ , optionally substituted by halogen, trifluoromethyl, hydroxyl, C _1-6 alkyl, amino or alkoxyl Represents any of the groups -CH = CH-CH = CH-;
R ₄ and R ′ ₄ are, independently of each other, H and a group selected from the group consisting of C _1-6 alkyl optionally substituted by halogen, trifluoromethyl, hydroxyl, amino or alkoxyl; Or each independently with R ₃ and R ′ ₃ independently of each other —CH═CH—CH═CH— group optionally substituted by halogen, trifluoromethyl, hydroxyl, C _1-6 alkyl, amino or alkoxyl Any one of; and
A is the following formula II

(Where m, n and p represent integers which can have the following values: m = 0, 1; n = 0, 1-10; p = 0, 1 where m, n and p are simultaneously 0. Never take the value of)))
Represents a spacer group having
A compound having R ₃ and R ₄ and R ′ ₃ and R ′ ₄ may be independently of each other, but together they may be optionally substituted by one or more halogen substituents —CH═CH—CH = CH- represents a group a compound according to claim 1. The following substituents:

The compound of claim 1 having The compound according to claim 3 , wherein the compound is 1,1 '-(biphenyl-4,4'-diylmethylene) bis [4- (4-chloro-N-methylanilino) quinolinium]. Q represents Br (bromide) or _F 6 P (hexafluorophosphate) The compound according to claim 3 and 4. a) Formula VII

[Where:
R ₁ represents a methyl group;
R ₂ represents a phenyl group substituted by one or more halogen substituents ;
R ₃ is a group selected from the group consisting of H, halogen, trifluoromethyl, hydroxyl, amino, and C _1-6 alkyl optionally substituted with trifluoromethyl, hydroxyl or amino, or R 3 ₄ represents any of the —CH═CH—CH═CH— groups optionally substituted by halogen, trifluoromethyl, hydroxyl, C _1-6 alkyl, amino or alkoxyl;
R ₄ is a group selected from the group consisting of H and C _1-6 alkyl optionally substituted by halogen, trifluoromethyl, hydroxyl or amino, or together with R ₃ , optionally halogen, tri Represents a —CH═CH—CH═CH— group optionally substituted by fluoromethyl, hydroxyl, C _1-6 alkyl, amino or alkoxyl]
The corresponding heterocyclic derivative of
And the dihalogenated derivative AX ₂ (where X represents a halogen atom: Cl, Br or I, and A represents

Represents
Or b) the corresponding heterocyclic derivative of formula VII and the dihalogenated derivative AX ₂ (where X is a halogen atom: Cl, Br or I). Is represented in an organic solvent in a 1: 1 molar ratio to give the mono-quaternized product, which is the first organic used in the reaction of AX ₂ with the first heterocyclic derivative of formula VII. The method for producing a compound according to claim 1, comprising reacting again with another different heterocyclic derivative molecule at a molar ratio of 1: 1 using an organic solvent having a polarity higher than that of the solvent. Formula: 4- (4-Chloro-N-methylanilino) quinoline

And 7-chloro-4- (4-chloro-N-methylanilino) quinoline

Having a compound.